MDA5 effect during HRV and RSV infection in respiratory epithelial cells.

A549 cells that had been transfected 48 hours earlier with siRNA to MDA5 or non-specific negative control siRNA, were uninfected or infected for 6, 12, 24, and 48 hours with HRV-B14 or RSV. For each time point, infections were performed in triplicate. Total cellular RNA were isolated using RNeasy Mini Kit (Qiagen). Multiplexed RNA libraries were prepared using the Truseq RNA sample prep kit (Illumina). Libraries were sequenced on a HiSeq 2000 Sequencing System (Illumina) to produce 50 bp single end reads. Sequencing reads were aligned with ELAND to the human reference genome version hg19.